Introduction

Experimental Design

Bone marrow-derived macrophages were obtained from tibias and femurs of mice as per protocols PP0000004702 (Harvesting and Plating Thioglycollate Elicited Macrophages for LM Distribution, pdf) and PP0000004602 (ATP / Kdo2-Lipid A treatment, pdf).
Cell treatments consisted of a) Control (no treatment); b) Kdo2-Lipid A pretreatment only (4hr pretreamemt,
no ATP); c) ATP only (with no Kdo2-Lipid A pretreatment); d) Kdo2-Lipid A pretreatment (4 hrs), followed
by addition of ATP at t=0. Lipid/mRNA measurements were taken at t=0 (control only), 15m, 30m, 1h, 2h, 4h, 8h, and 20h
(relative to ATP addition at t=0). Measurements were normalized to the average DNA value across all timepoints for each
treatment and expressed as pmol analyte/ug DNA.